Аннотация:FACT (facilitates chromatin transcription) is a histone chaperonethat participates in nucleosome removal and reassembly duringtranscription and replication. Previously we have shown thatyeast FACT dramatically alters the nucleosome structure withoutATP hydrolysis, but the extent of these alterations depends onthe presence of HMGB domain-containing protein Nhp6. Thedetailed mechanism of this process remains unknown. HereFACT, FACT:Nhp6 and FACT:Nhp6: nucleosome complexeswere studied using electron microscopy. All complexes are highlyflexible and adopt broad ranges of configurations. DNA-bindingprotein Nhp6 binds to the C-terminal tails of both FACT subunits and induces formation of more open FACT complexes,thus altering the structures of FACT and the nucleosomes and facilitating nucleosome unfolding. The data suggest that Nhp6 has multiple roles before and during nucleosome unfolding byFACT, and that the process proceeds through a series of energetically similar intermediate structures, ultimately leading to anextensively unfolded form. We proposed FACT-dependent nucleosome unfolding pathway based on a large number of potentialintermediates revealed by electron microscopy. This work wassupported by the Russian Science Foundation (#19-74-30003).Electron microscopy was performed on the Unique equipmentsetup “3D-EMС” of Moscow State University, Department of Biology.